Figure S5.
Analysis of dynamin-deficient olf hair cells. (A) Quantification of PMI volume and depth measured from FIB-SEM reconstructions of three olf hair cells of y w, gox, and shits2/Y pupae. Data are shown as box-and-whisker plots indicating the median, interquartile range, and full distribution. Statistical significance was determined using the Mann–Whitney U test (two-sided). **: P < 0.01; n.s.: not significant. (B) Phenotype of maxillary palp bristles. Female (shits2/X, control) and male (shits2/Y, experimental) progenies from the same batch of heat-treated flies are compared. Filled dot indicate olf bristles of normal length and abnormal morphology, respectively. Arrowhead in the shits2/Y panel indicates the open hole of the cuticle. (C) Enlargement of abnormal olf bristles of shits2/Y. Left to right: short, branched, and hole phenotype. (D) FIB-SEM views of olf hair cells #2 and #3 of shits2/Y showing ER, plasma membrane, and their contact site (yellow dot). (E) Maxillary palp expressing UAS-shits1 driven by the neur-Gal4 driver. Apical ECM marker: mVenus-Tyn and mScarlet-Dyl. Arrowhead indicates the lumen of invaginated olf cells containing secreted mVenus-Tyn and surrounded by mScarlet-Dyl. Enlargement shows a hair cell–like remnant of Tyn left outside of the invaginated olf cell. (F) Number of olf hairs on the maxillary palp of shits2 mutants and gox knockdown combinations. (G) Number of pores of maxillary palp in the same genotypes. Each data point represents one maxillary palp (n = 17 for shits2/X; n = 25 for shits2/Y; n = 18 for shits2/X; gox KD; n = 22 for shits2/Y; gox KD). Data are shown as mean ± SD, and statistical significance was determined using the Mann–Whitney U test (two-sided). **: P < 0.01; n.s.: not significant. Note that neur > gox RNAi partially suppressed the shts2/Y phenotype. (H) shi-GFP expression driven by the neur-Gal4 driver in the olf and mech at 42 h APF. GFP signal is highly enriched in the cortical region of the olf hair cell, but its intensity is much lower in the mech. Socket cells of olf and mech accumulate about the same levels of GFP signal. (I) PLCdelta-PH-EGFP, a marker for PI(4,5)P₂, driven by the neur-Gal4 driver. EGFP signal strongly labels the cortical region of the olf hair cell and weakly that of the mech hair cell. Bar: 20 µm (B), 1 µm (C) 1 µm (D), and 10 µm (E, H, and I). Asterisk: **, P < 0.001 (A and F); (G). Statistical significance was determined by Mann–Whitney U test.

Analysis of dynamin-deficient olf hair cells. (A) Quantification of PMI volume and depth measured from FIB-SEM reconstructions of three olf hair cells of y w, gox, and shits2/Y pupae. Data are shown as box-and-whisker plots indicating the median, interquartile range, and full distribution. Statistical significance was determined using the Mann–Whitney U test (two-sided). **: P < 0.01; n.s.: not significant. (B) Phenotype of maxillary palp bristles. Female (shits2/X, control) and male (shits2/Y, experimental) progenies from the same batch of heat-treated flies are compared. Filled dot indicate olf bristles of normal length and abnormal morphology, respectively. Arrowhead in the shits2/Y panel indicates the open hole of the cuticle. (C) Enlargement of abnormal olf bristles of shits2/Y. Left to right: short, branched, and hole phenotype. (D) FIB-SEM views of olf hair cells #2 and #3 of shits2/Y showing ER, plasma membrane, and their contact site (yellow dot). (E) Maxillary palp expressing UAS-shits1 driven by the neur-Gal4 driver. Apical ECM marker: mVenus-Tyn and mScarlet-Dyl. Arrowhead indicates the lumen of invaginated olf cells containing secreted mVenus-Tyn and surrounded by mScarlet-Dyl. Enlargement shows a hair cell–like remnant of Tyn left outside of the invaginated olf cell. (F) Number of olf hairs on the maxillary palp of shits2 mutants and gox knockdown combinations. (G) Number of pores of maxillary palp in the same genotypes. Each data point represents one maxillary palp (n = 17 for shits2/X; n = 25 for shits2/Y; n = 18 for shits2/X; gox KD; n = 22 for shits2/Y; gox KD). Data are shown as mean ± SD, and statistical significance was determined using the Mann–Whitney U test (two-sided). **: P < 0.01; n.s.: not significant. Note that neur > gox RNAi partially suppressed the shts2/Y phenotype. (H) shi-GFP expression driven by the neur-Gal4 driver in the olf and mech at 42 h APF. GFP signal is highly enriched in the cortical region of the olf hair cell, but its intensity is much lower in the mech. Socket cells of olf and mech accumulate about the same levels of GFP signal. (I) PLCdelta-PH-EGFP, a marker for PI(4,5)P₂, driven by the neur-Gal4 driver. EGFP signal strongly labels the cortical region of the olf hair cell and weakly that of the mech hair cell. Bar: 20 µm (B), 1 µm (C) 1 µm (D), and 10 µm (E, H, and I). Asterisk: **, P < 0.001 (A and F); (G). Statistical significance was determined by Mann–Whitney U test.

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